臺灣博碩士論文加值系統

稻熱病由Magnaporthe oryzae引起並造成嚴重危害，由於田間生理小種變異快速，導致水稻抗性不易維持，為有效減少病害發生，必須持續選育抗病品種。為延續實驗室先前成果，以11個分別具廣幅抗性之IRBLs為抗性貢獻親本，透過分子標誌輔助回交育種法 (marker-assisted backcrossing) 將抗病基因分別導入高雄145號。本研究運用前人設計之分子標誌，或以tetra-primer amplification refractory mutation system-polymerase chain reaction (T-ARMS-PCR) 技術新開發之分子標誌，針對不同雜交組合，於各個世代進行抗病基因型檢定，並選用目標R genes對其呈現抗病之菌株進行人工接種抗性檢定，根據檢定結果及田間農藝性狀觀察，選取抗病且性狀固定之品系。經持續選拔後，獲得 R genes 為同型合子之子代，進入初級或高級產量比較試驗，並以單植或混植法於嘉義農業試驗分所或臺東關山病圃進行抗性檢定。最後以genotyping-by-sequencing (GBS) 進行背景基因型選拔，選出與高雄145號遺傳背景相似度高且抗病基因型已固定之品系。目前三個雜交組合篩選至BC3F6、六個雜交組合至BC3F5、一個雜交組合至BC3F4與一個雜交組合至BC4F4，背景回復率78.1-97.6%。其中推進最快之「高雄145號×IRBLta2-Pi」、「高雄145號×IRBLkh-K3」與「高雄145號×IRBL9-W」三個雜交組合，已篩選出10個呈現顯著抗病性、且農藝性狀及米質均與高雄145號相近之優良品系。未來新品種育成後，可望以單一品種種植或混合種植方式，於田間達到持久抗性之效果。

Rice blast can cause serious yield loss in rice production. Due to rapid evolution of Magnaporthe oryzae population, rice resistance to blast is generally easy to be overcome. To effectively control the disease, it is important to keep developing resistant varieties. As part of the ongoing effort in our lab, this study continues to introgress resistance genes from 11 IRBLs into Kaohsiung 145 through marker-assisted backcrossing. The individuals were screened for R genes by previously designed primers and new primers developed based on tetra-primer amplification refractory mutation system-polymerase chain reaction (T-ARMS-PCR) method. They were also evaluated for blast resistance by artificial inoculation with incompatible M. oryzae isolates, then selected according to their stable resistance and agronomic performance in the field. The progeny carried homozygous R genes were tested for agronomic traits in preliminary or advanced yield trials, and evaluated for resistance in Chiayi and Taitung blast nursery fields. The lines with high recovery rate of the recurrent Kaohsiung 145 genome and resistance allele in the homozygous state were selected by genotype-by-sequencing (GBS). To date, 3, 6, 1, and 1 sets of the “Kaohsiung 145 × IRBL” BC3F6, BC3F5, BC3F4, and BC4F4 have been generated, respectively, with background recovery rates ranging from 78.1 to 97.6 %. Ten elite lines from KH145×IRBLta2-Pi, KH145×IRBLkh-K3 and KH145×IRBL9-W showed moderate to high blast resistance in artificial inoculation test and blast nursery field. Furthermore, the agronomic traits and rice quality of them were similar to KH145. The deployment of KH145 multiline varieties will help achieve stable and long-term control of rice blast.

誌謝 I
中文摘要 II
Abstract III
目錄 IV
表目錄 VI
圖目錄 VII
前言 1
Chapter 1 前人研究 3
1.1 水稻抗稻熱病基因 3
1.2 稻熱病判別品系 4
1.3 分子標誌輔助育種 4
1.4 高通量基因型分析 5
1.5 持久抗病 6
1.6 親本來源與特性 6
Chapter 2 材料方法 8
2.1 族群建立 8
2.2 水稻DNA萃取 9
2.3 水稻抗稻熱病基因座之定序 9
2.4 R gene前景基因型篩選 10
2.4.1 分子標誌設計 10
2.4.2 PCR反應與電泳分析 10
2.5 背景基因型分析 11
2.5.1 Genotyping by sequencing (GBS) 11
2.5.2 GBS資料與分析 12
2.5.3 背景基因型回復率分析 13
2.6 抗性檢定 13
2.6.1 水稻栽種 13
2.6.2 供試稻熱病菌、培養條件與接種源製備 14
2.6.3 人工接種抗性檢定 14
2.6.4 稻熱病圃抗性檢定 15
2.7 初級產量比較試驗 16
Chapter 3 結果 18
3.1 抗病基因座之定序分析 18
3.2 R gene分子標誌測試 18
3.3 11個IRBLs與高雄145號雜交子代分析 19
3.3.1 2016年第二期作 19
3.3.2 2017年第一期作 19
3.3.3 2017年第二期作 19
3.3.4 2018年第一期作 20
3.3.5 2018年第二期作 20
3.3.6 2019年第一期作 20
3.4 人工接種抗性檢定 20
3.5 病圃抗性檢定 21
3.6 初級產量比較試驗 22
3.7 背景基因型分析 22
Chapter 4 討論 24
參考文獻 30
附表 37
附圖 62
附錄 85

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